Penicillin antibiotics

ABSTRACT

WHERE R is a hydrogen atom or an organic group and Ra is a hydrogen atom or an acyl group. The compounds are syn isomers or exist as mixtures containing at least 75% of the syn isomer. These antibiotic compounds possess high antibacterial activity against a range of gram positive and gram negative organisms coupled with particularly high stability to Beta -lactamases produced by various gram negative organisms. The invention is also concerned with the administration of the compounds.   The invention provides novel antibiotic compounds which are 6 Beta -acylamidopenam-3-carboxylic acids and non-toxic derivatives thereof, characterized in that the acylamido group has the structure

United States Patent [1 1 Gregson et al.

[ PENICILLIN ANTIBIOTICS [75] Inventors: Michael Gregson, London; Martin Christopher Cook; Gordon Ian Gregory, both of Chalfont St. Peter, all of England [73] Assignee: Glaxo Laboratories Limited,

Greenford, England 221 Filed: May 3, 1974 211 App]. No.: 466,780

Related US. Application Data [60] Division of Ser. No. 274.602, July 24, I972, which is a continuation-in-part of Ser. No. 22l,057, Jan. 26, I972, abandoned.

[52] US. Cl 260/239.1; 424/271 [5i] Int. Cl. C07D 499/44 [58] Field of Search 260/2391 [56] References Cited FOREIGN PATENTS OR APPLICATIONS l,l59,449 I963 Germany 260/2391 OTHER PUBLICATIONS Arkiv for Kemi, 22 (33), PP. 451-467, (1964).

Acta Chemica Scandinavica, l9, pp. 281-299, (I965). Chemical Abstr., Vol. 70, Col. 77850(c), (1969). Acta Poloniae Pharmaceutica, I968, 25(5), pp. 507-511.

[4 1 Dec. 16, 1975 Primary Examiner-Gerald A. Schwartz Attorney, Agent, or Firm-Bacon & Thomas ABSTRACT The invention provides novel antibiotic compounds which are 6/3-acylamidopenam-3-carboxylic acids and non-toxic derivatives thereof, characterized in that the acylamido group has the structure R.C.CONH

2 Claims, No Drawings PENICILLIN ANTIBIOTICS CROSS REFERENCE TO RELATED APPLICATION This application is a division of pending application Ser. No. 274.602 filed July 24. 1972 which in turn is a continuation-in-part of application Ser. No. 22l.()57 filed Jan. 26. [972 and now abandoned.

This invention is concerned with improvements in or relating to antibiotics of the penicillin series.

The penicillin compounds referred to in this specification are generally named with reference to penam (J. Amer. Chem. Soc. 1953, 75. 3293).

As is well known. antibiotics of the penicillin series are 6B-acylamidopenam-S-carboxylic acids and their various non-toxic derivatives e.g. salts. esters, amides. hydrates or the corresponding sulphoxides. Such antibiotics may. for example. carry substituents on at least one of the gem-dimethyl groups normally present at the 2-position.

The new compounds of the present invention are characterized in that said acylamido group of the penicillin antibiotic in an oz-hydroxyiminoacylamido or aacyloxyiminoacylamido group. the compounds being syn isomers or mixtures wherein the syn isomeric form predominates.

According to one embodiment of the invention. therefore. we provide compounds selected from the group consisting of o B-acylamidopenam-B-carboxylic acids and nontoxic derivatives thereof characterized in that said acylamido group has the structure:

where R is a hydrogen atom or an organic group and R" is a hydrogen atom or an acyl group, said compounds being syn isomers or existing as mixtures containing at least 757: of the syn isomer.

'The compounds of the invention are defined as having the syn (cis) isomeric form as regards the configuration of the group OR" with-respect to the carboxamido group. Preferably. the mixtures of isomers contain at least 90% of the syn isomer and not more than l7r of the anti isomer.

In this specification. the syn configuration is structurally denoted thus:

and the anti configuration thus:

These configurations are allocated on the basis of the work of Ahmad and Spencer. Can. J. Chem. l96l. 39. I340.

2 The compounds of the invention may be defined l the formula:

(:0 Ml B /CH3 R.C. Tl-

1 ca. O N i (3 where R and. R" have the above-defined meanings an B is S or S 0. preferably 5. The invention als includes compounds not specifically embraced by to mula (l). e.g. ZB-acetoxymethyl pcnicillins.

The term non-toxic as applied to the derivatives the compounds of the invention means those deriva lives which are physiologically acceptable in the dosag at which they are administered.

Salts which may be formed. where applicable. fro: the compounds according to the invention include inorganic base salts such as alkali metal. cg. sodiui and potassium. alkaline earth metal e.g. calcium. an organic base. e.g. procaine. phenylethylbenzylamin and dibenzylethylene diamine. salts and (h) acid add tion salts e.g., with hydrochloric. hydrobromic. su phuric. nitric. phosphoric. toluenep-sulphomc an methane sulphonic acids. The salts may also be in th form of resinates. formed e.g.. with a polystyrene resi containing amino. quaternary amino. or sulphonic aci groups. or a resin containing carboxyl groups. eg polyacrylic acid resin. The resin may if desired r cross-linked. c.g. it may be a copolymer of styrene an divinyl-benzene containing the appropriate group Additionally. the derivatives may be in the form of chelate with a heavy metal such iron or copper.

The compounds of the invention. including the not toxic derivatives thereof. are characterized by the high antibacterial activity against a range of gram-pos tive and gram-negative organisms coupled with partici lary high stability of B-lactamases produced by varioL gram negative organisms.

The group R" in the above formulae. when it is acy may be chosen from a wide variety of possible group Thus the group R" may be a carboxylic acyl grou RCO having from l to 20 carbon atoms. In particula R may be an aliphatic. cycloaliphatic or aromati group or it may be such an organic group linked to ti carbonyl group through an oxygen atom. a sulphi atom or an imino group. Such an aliphatic. cycloal phatic or aromatic group may be substituted by halt gen (F. Cl, Br or I). amino. nitro. C C alkyl. C,( alkoxy etc.

Particular examples of R" include alkanoyl. alkenoy and alkynoyl of up to 7 carbon atoms c.g. acetyl. prop onyl. butyryl. acrylyl. crotonyl; substituted. e.g. halog' nated (F. Cl. Br. I) or amino or substituted amino d1 rivatives of such groups eg chloroacetyl. dichloro: cetyl or B-aminopropionyl; alkoxycarbonyl of up to carbon atoms e.g. ethoxycarbonyl. and t-butoxycarbr nyl; substituted alkoxycarbonyl e.g. 2.2.2-trichloro| 3 thoxycarbonyl; alkylthiocarbonyl of up to 7 carbon atoms and substituted derivatives thereof; aralkyloxycarbonyl e.g. benzhydryloxycarbonyl and benzyloxycarbonyl'. C -C aroyl e.g. benzoy] and substituted. e.g. nitrated derivatives of such groups e.g. nitrobenroyl; and substituted or unsubstituted carbamoyl or thiocarbamoyl i.e. compounds in which the group R" has the formula (R") N.CO or (R") N.CS and R" is the same or different and each is a hydrogen atom or a substituent such as alkyl of 1-7 carbon atoms e.g. ethyl or methyl and substituted. e.g. halogenated. alkyl of l7 carbon atoms e.g. chloroethyl, or R". where R has the meaning defined below.

The group R in the above general formulae may be chosen from the following list which is not intended to be exhaustive:

i. Hydrogen.

ii. R". where R" is aryl (carbocyclic or heterocyclic). cycloalkyl. substituted aryl. substituted cycloalky]. cycloalkadienyl, or a non-aromatic or mesionic group. Examples of this group include phenyl; naphthy]: phenyl or naphthyl substituted by halo e.g. chloro or bromo. hydroxy. lower alkyl e.g. methyl, nitro. amino. lower alkylamino e.g. methylamino. diloweralkyl amino e.g. dimethylamino. lower all-tanoyl e.g. acetyl. lower alkanoylamido. lower alkoxy e.g. methoxy or ethoxy. or lower alkylthio e.g. methylthio; a S or 6- membered heterocyclic group containing at least one hetero atom selected from S. N and O e.g. thien-Z-yl. thien-3-yl. furyl. pyridyl. 3- or 4-isoxazolyl; substituted 3- or J-isoxazolyl e.g. 3-aryl-5-methylisoxazol-4-yl. the aryl group being e.g. phenyl or halophenyl; cyclohexyl; cyclopentyl; sydnone; and cyclohexadienyl.

iii. R"(CH J,,,Q,,(CH where R" has the above defined meaning and m is O or an integer from 1 to 4.11 is 0 or I. p is an integer from 1 to 4 and Q is S. O or NR wherein R is hydrogen or an organic group e.g. alkyl such as methyl or aryl such as phenyl. Examples of this group include methyl. ethyl or butyl substituted by the various specific R" groups listed under (ii) e.g. benzyl and the appropriate substituted benzyl groups.

iv. C,,H ,,s wherein n is an integer from I to 7. The group may be straight or branched and, if desired. may be interrupted by an oxygen or sulphur atom or the group NR wherein R is hydrogen or an organic group e.g. alkyl such as methyl or aryl such as phenyl; or be substituted by a cyano. carboxy. alkoxycarbonyl, hydroxy or carboxycarbonyl (HOOCCO) group or by a halogen atom. Examples of such groups include hexyl.

heptyl. butylthiomethyl. cyanomethyl or trihalomethyl.

Preparation The compounds according to the invention may be prepared by any convenient method. According to one embodiment of the invention we provide a process for the preparation of a compound of the formula CH R.C.CO.NH:[( B 3 H CH3 N 0 N (wherein R is a hydrogen atom or an organic group. R" is hydrogen or an acyl group and B is S or S O) and derivatives thereofl which comprises either (A) condensing a compound of the formula B ca a N 3 N cs 4-- 0 cooa (wherein B has the above defined meaning and R is hydrogen or a carboxyl blocking group) with an acylating agent. advantageously the syn isomer. corresponding to the acid the resulting precursor acyl group into the desired acyl group; or (B) reacting a compound of the formula (Illl B 3 O=C==N N (IV) 0 1 COOR (wherein B and R have the above defined meaning except that R is not hydrogen) with an acid of formula (III) wherein R" is not hydrogen; whereafter. if necessary and desired in each instance. any of the following reactions (C) are carried out (i) conversion of a precursor for the desired R.C.COOH

where R and R" have the above defined meanings, with an amino compound H N B CH3 cooa I) [where B has the above defined meaning and R is hydrogen or a carboxyl blocking group e.g. the residue of an ester-forming alcohol (aliphatic or araliphatic), phenol. silanol. stannanol or acid] the condensation, if desired. being effected in the presence of a condensation agent. and being followed, if necessary, by removal of the group R. There may also be used a derivative of the amino compounds such as a salt e.g. a tosylate.

Compounds in which R" is hydrogen may be prepared indirectly and compounds in which R" is not hydrogen may be prepared directly by employing as|the acylating agent an acid halide, particularly an acid chloride or bromide. In the preparation of a compound in which R" is hydrogen by this technique an additional step will be necessary in which the acyl group R" is removed to yield the hydroxyimino compound. The acylation may be effected at temperatures of from 50 to +50C, preferably from to +20C e.g. about 0C. The acylating agent may be prepared by reacting the acid (Ill) in which R is not hydrogen with a halogenating agent e.g. phosphorus pentachloride, thionyl chloride or oxalyl chloride. The acylation may be effected in aqueous or non-aqueous media and suitable media include an aqueous ketone such as aqueous acetone, an ester e.g. ethyl acetate, or an amide e.g.

6 dimethylacetamide. or a nitrile e.g. acetonitrile. L mixtures thereof.

Acylation with an acid halide may be effected in th presence of an acid binding agent e.g. a tertiary amin (e.g. triethylamine or dimethylaniline). an inorgani base (e.g. calcium carbonate or sodium bicarbonate or an oxirane, which binds hydrogen halide liberated i the acylation reaction. The oxirane is preferably a lov enLZ-alkylene oxide e.g. ethylene oxide or propylen oxide.

When using the free acid form of a compound formula (Ill) and. if desired. where R"=H, suitabl condensing agents for use in the preparation of tt compounds according to the invention include ca bodiimides, for example N.N'-diethyl', dipropyldiisopropylcarbodiimide. N,N-dicyclohexylcarbodi mide, or N-ethyl-N-'y-dimethylaminopropylcarbodi mide; a suitable carbonyl compound, for example ca bonyldiimidazole; or an isoxazolinium salt, for exan ple. N-ethyl-S-phenylisoxazolinium-3-sulphonate an N-t-butyl-5-methylisoxazolinium perchlorate. The cor densation reaction is desirably effected in an anhydroi reaction medium. e.g. methylene chloride. dimethy formamide or acetonitrile. since one may then regular more precisely reaction parameters such as temper; ture.

Alternatively. acylation may be effected with 0th amide-forming derivatives of the free acid such as. f( example, a symmetrical anhydride or mixed anhydrid eg. with pivalic acid or formed with a haloformate, e. a lower alkylhaloformate. The mixed or symmetric anhydrides may be generated in situ. For example. mixed anhydride may be generated using N-ethoxyca bonyl-2-ethoxy-l, 2-dihydroquinoline. Mixed anh drides may also be formed with phosphorus acids (ft example phosphoric or phosphorous acids), sulphur acid or aliphatic or aromatic sulphonic acids (for exan ple p-toluene sulphonic acid). Another conveniei acylating agent is an activated ester e.g. a compound the formula where W, is for example. azide. oxysuccinimide. o ybenztriazole, pentachlorophenoxy or p-nitrophenox One may prepare compounds according to the inve tion in which R" H by condensing an acylating age corresponding to the acid (III) in which R" is not h drogen but is a group which can be readily removed yield the desired group =N-OH with a compound formula ll. The R" group is removed subsequently. desired in conjunction with the removal of the grol R. illustrative of such readily removable R" groups a acetyl, if desired having at least one electron-withdra ing group on the a-carbon atom e.g. trichloroacet dichloroacetyl, monochloroacetyl, trifluoroacetyl, r fluoroacetyl and monofluoroacetyl; formyl; benzh dryloxycarbonyl. benzyloxycarbonyl. t-butoxycarbor and 2,2.2-trichloroethoxycarbonyl. The removal such groups may for example be effected under milc basic conditions. Thus. for example. an acetyl gro may be removed by means of treatment with aqueo alkali. Halogenated acetyl groups may be removed means of aqueous bicarbonate and. additionally. chl

roacctyl can be removed using a nucleophilc such as a thiourea. Ben7hydryloxycarbonyl and t-butovycarbonyl groups can be removed using trifluoroacetic acid with or without anisole. 2.2l-Trichloroethoxycarbonyl may be removed by means of a reducing agent such as zinc/acetic acid or zinc/formic acid. It will be appreci ated that although the preparation of compounds hav ing readily remo\ able R" groups affords a convenient route to hydrosyimino compounds, compounds with such R" groups are also compounds according to the invention and may possess desirable properties in their own rightv Alternatively the compound of formula (I) may be prepared from a compound of formula COOR V) where B and R have the above defined meanings (except that R is not H) by reaction with an acid of formula (lll) (where R" is not H) and subsequently removing the groups R and. if desired. R (see for example Dutch Patent Application No. 6808622).

The reaction of the compound of formula (ll) or (IV) may be carried out towards the end of the preparative sequence. the only additional reactions being deprotection reactions and purifications.

Compounds of formula I where R" acyl may be obtained from the corresponding compounds of formula l where R" H or from esters thereof (i.e. having at the 3-position a group COOR. R having the above defined meaning) by acylation. Acylation may be ef fected in any convenient manner cg. using an acid halide. symmetrical or mixed anhydride. ketene. acyl azide or carbodiimide (when the 3-carboxy group is protected) corresponding to the acid R"OH. Alterna tively the acylation may be effected by means of a haloformate for example a chloroformate such as ethylchloroforrnate whereupon one will obtain carbonates or by means of an isocyanate R"NCO for example 2- chloroethyl isocyanate whereupon one will obtain carbamates in which OR" has the formula R"NHC0.0 where R" has the above defined meaning. The acylation may be catalysed e.g. by a base such as triethylamine. diethylaniline. pyridine. propylene oxide. magnesium oxide. sodium carbonate or calcium carbonate. The acylation may be effected in an organic solvent. Suitable solvents include halogenated hydrocarbons e.g. methylene chloride; cyclic ethers eg. dioxan or tetrahydrofuran; nitriles e.g. acetonitrile; nitrohydrocarbons eg. nitromethane; esters e.g. ethyl acetate; or the acylating agent itself. The acylation may be effected at a temperature of -l() to +100C. preferably to 50C advantageously 0 to C. After the acylation has been effected the group R is removed. if necessary.

if desired. one can first prepare a compound of for mula R.CO.CO.NH K 3 N 3 0 1 (vi) COOR (where R. R and B have the above defined meanings) and then effect reaction of the compound of formula (Vi) \vith R"O.NH. (R" having the above defined meaning). followed. if necessary by removal of the group Rv The reaction product may be separated into syn and anti isomers before or after removal of R.

One may prepare compounds of formula (I) herein R is an activating group such as cyano or pyridyl by a technique invol ing nitrosation. Thus a compound possessing the acylamido group OOH may be nitrosated using. for example. nitrous acid (which may be prepared in situ by reaction of an alkali metal nitrite with a weak acid e.g. acetic acid), nitrosyl chloride. or an organic nitrosating agent e.g. an alkyl, cycloalkyl. aryl or aralkyl nitrite. In the case of nitrosa tion of a compound containing the group concomitant decarboxylation will occur. Separation of syn and anti-isomers may be necessary after the nitro sation reaction.

Compounds of the formula (ll) may be employed as esters; those of formula (IV) are esters. One may also use the free amino acid or an acid addition salt of the free amino acid or ester thereof. Salts which may be used include acid addition salts e.g. with hydrochloric. hydrobromic, sulphuric. nitric. phosphoric. toluene-p sulphonic and methane sulphonie acids.

The ester may be formed with an alcohol, phenol, silanol or stannanol having up to 20 carbon atoms which may readily be split off at a later stage of the overall reaction.

Any esterifying group substituting the 4-carboxyl group of a compound of formula (ll). (IV) or (VI) is preferably formed with an alcohol (aliphatic or araliphatic). phenol. silanol, stannanol or acid which may readily be split off at a later stage of the reaction.

Suitable esters thus include compounds containing as ester group a group selected from the following list which is not intended to be an exhaustive list of possible ester groups.

i. COOCR"R"R" wherein at least one of R. R" and R" is an electrondonor eg p-methoxyphenyl. 2.4.6- trimethylphenyl. J-anthryl, methoxy. acetoxy, or fur- 2yl. The remaining R". R" and R" groups may be hydrogen or organic substituting groups. Suitable ester groups of this type include p-methoxybenzyloxycarbo- 9 nyl and 2.4.o-trimethylbenzyloxy carbonyl.

ii. COOCR"R"R wherein at least one of R". R and R" is an electron-attracting group e.g. benzoyl. p-nitrophenyl. 4-pyridyl. trichloromethyl. tri' bromomethyl. iodomethyl. cyanomethyl. ethoxycarbonylmethyl. arylsulphonylmethyl. Z-dimethylsulphoniumethyl. o-nitrophenyl or cyano. The remaining R. R". and R" groups may be hydrogen or organic substituting groups. Suitable esters of this type include benzoylmethoxycarbonyl. p-nitrobenzyloxycarbonyl. 4 pyridylmethoxycarbonyl. 2.2.Z-trichloroethoxycarbonyl and 2.2.2-tribromoethoxycarbonyl.

iii. CO()CR"R"R wherein at least two of R". R" and R" are hydrocarbon such as aikyl e.g. methyl or ethyl. or aryl e.g. phenyl and the remaining R". R" and R group. if there is one. is hydrogen. Suitable esters of this type include t-butyloxycarbonyl. tamyloxycarbonyl. diphenylmethoxycarbonyi and triphenylmethoxycarbonyl.

iv. COOR" wherein R is adamantyl. 2-benzyloxyphenyl. 4methylthiophenyl. tetrahydrofur-Z-yl or tetrahydropyran-2-yl.

v. Silyloxycarbonyl groups obtained by reaction of a carboxyl group with a derivative ofa silanol. The derivative of a silanol is conveniently a halosilane or a silazane of the formula RHSiX; R SiX RfiSiNRE; RfiSiNHSiRy. R -.Si.NH.COR; RfiSiNHCONH- .SiR*=,; RNHCONRSiRk; or RC(OSiR" NSiR' where X is a halogen and the various groups R. which can be the same or different. represent hydrogen atoms or alkyl. e.g. methyl. ethyl. n-propyl. iso-propyl; aryl. e.g. phenyl; or aralkyl e.g. benzyl groups. Preferred derivatives of silanols are silyl chlorides such as for example trimethylchlorosilane and dimethyldichlorosilane.

The carboxyl group may be regenerated from an ester by any of the usual methods. for example. acidand base-catalysed hydrolysis is generally applicable. as well as enzymically-catalysed hydrolyses; however. aqueous mixtures may be poor solvents for these compounds and they may cause isomerizations. rearrangements. side-reactions. and general destruction. so that special methods may be desirable.

Five suitable methods of deesterification are 1. Reactions with Lewis acids.

Suitable Lewis acids for reaction with the esters include trifluoroacetic acid, formic acid, hydrochloric acid in acetic acid. zinc bromide in benzene and aqueous solutions or suspensions of mercuric compounds. The reaction with the Lewis acid may be facilitated by addition of a nucleophile such as anisole.

2. Reduction.

Suitable systems for effecting reduction are zinc/acetic acid. zinc/formic acid. zinc/lower alcohol. zinc- /pyridine. palladised-charcoal and hydrogen. and sodium and liquid ammonia.

3. Attack by nucleophiles.

Suitable nucleophiles are those containing a nucleophilic oxygen or sulphur atom for example alcohols. mercaptans and water.

4. Oxidative methods. for example. those which involve the use of hydrogen peroxide and acetic acid.

5. Irradiation.

Where at the end of a given preparative sequence compounds are obtained wherein B is S O and a compound is desired in which B is S conversion to a sulphide may for example. be effected by reduction of the corresponding acyloxysulphonium or alkyloxysulphonium salt prepared in situ by reaction with e.g. acetyl chloride in the case of an acctoxysulphonium salt. reduction being effected by. for example. sodium dithionite or by iodide ion as in a solution of potassium iodide in a water miscible solvent e.g. acetic acid. tetra hydrofuran. dioxan. dimethylformamitle or dimethylacetamide. The reaction may be effected at a temperature of2() to +5()C.

Alternatively. reduction of the l-sulphinyl group may be effected by phosphorus trichloride or trihromide in solvents such as methylene chloride. dimethylforrnam- 'ide or tetrahydrofuran, preferably at a temperature of 20C to +50C The acid (ill) to which the acylating agent corresponds may be obtained by reacting the glyoxylic acid.

R.C().(O()H (where R has the above defined meaning) or an ester thereof with R"O.NH. (R" having the above defined meaning).

The resulting acid or ester may then be separated into its syn and anti isomers e.g. by crystallisation. chromatography or distillation. followed when necessary by hydrolysis of the ester.

Alternatively the acid (Ill) where "=H may be ob tained by reacting an ester of the acid R.CH- .(OOH (where R has the above defined meaning) with an inorganic or organic nitrosating agent e.g. an alkyl nitrite such as isopropyl nitrite. or nitrosyl chloride in the presence of an acid or base. We prefer that an excess of the nitrosating agent be used e.g. a molar excess. Thereafter the ester group is removed. if necessary.

Syn and anti isomers may be distinguished by appropriate techniques. eg by their ultraviolet spectra. by thin layer or paper chromatography or by their nuclear magnetic resonance spectra. For example. for DMSO- d solution compounds of Formula I exhibit the doublet for the amide NH at a lower field for the syn isomers than for the anti-isomers). These factors may be em ployed in monitoring reactions.

The antibacterial compounds according to the invention may be formulated for administration in any con venient way. by analogy with other antibiotics and the invention therefore includes within its scope a pharmaceutical composition comprising an antibacterial compound of formula I or a non-toxic derivative e.g. salt thereof (as herein defined) adapted for use in human or veterinary medicine. Such compositions may be presented for use in conventional manner with the aid of any necessary pharmaceutical carriers or excipient.

The antibacterial compounds according to the invention may be formulated for injection and may be presented in unit dose form in ampoules. or in multidose containers with an added preservative. The compositions may take such forms as suspensions. solutions. emulsions in oily or aqueous vehiclels. and may contain formulatory agents such as suspending. stabilising and- ]or dispersing agents. Alternatively the active ingredient may be in powder form for reconstitution with a suitable vehicle. e.g. sterile. pyrogen-free water. before use.

The compositions may be presented in a form suitable for absorption by the gastrointestinal tract. Tablets and capsules for oral administration may be in unit dose presentation form. and may contain conventional excipients such as binding agents. for example. syrup. acacia. gelatin. sorbitol. tragacanth. or polyvinyl-pyrollidone; fillers. for example lactose. sugar. maize-starch.

calcium phosphate. sorbitol or glycine. lubricants, for example. magnesium stearate. talc. polyethylene glycol. silica: disintegrants. for example. potato starch or acceptable wetting agents such as sodium lauryl sulphate. The tablets may be coated according to methods well known in the art. Oral liquid preparation may be in the form of aqueous or oily suspensions, solutions. emulsions. syrups. elixirs. etc. or may be presented as a dry product. for reconstitution with water or other suitable vehicle before use. Such liquid preparations may contain conventional additives such as suspending agents. for evarnple. sorbitol syrup. methyl cellulose. glucoselsugar sy rup. gelatin. h droxyethylcellulose. carboxymcthyl cellulose. aluminium stearate gel or hydrogenated edible tats. emulsifying agents. for example. lecithin. sorbitan monoolcate or acacia; non-aqueous vehicles which may include edible oils. for exam ple. almond oil. fractionated coconut oil. oily esters. propylene glycol. or ethyl alcohol. preservatives. l'or e\amplc. methyl or propyl p-hydroxybenzoatcs or sorbic acid. Suppositories will contain conventional suppository bases. cg cocoa butter or other glyceride.

The composition may also be prepared in suitable forms for absorption through the mucous membranes of the nose and throat or bronchial tissues and may con\ eniently take the form of powder or liquid sprays or inhalants. lorenges. throat paints etc. For medication of the eyes or cars. the preparations may be pres ented as individual capsules. in liquid or semi-liquid form. or may be used as drops etc. Topical applications may be formulated in hydrophobic or hydrophilic bascs as ointments. creams. lotions. paints. powders etc.

For veterinary medicine the composition. may. for

example. be formulated as an intramammary preparation in either long acting or quick-release bases.

The compositions may contain from 0.1% upwards. preferably from 104100? of the active material. depending on the method of administration. Where the compositions comprise dosage units. each unit will preferably contain 50-500 mg. of the active ingredient. The dosage as employ ed for adult human treatment will preferably range from 100-3000 mg. for instance 1500 mg per day. depending on the route and frequency of administration.

The compounds according to the invention may be administered in combination with other therapeutic agents such as antibiotics. for example other cephalosporins. the penicillins or tetracyclines.

The following examples illustrate the invention.

2-HYDROXYIMINO'ACETIC ACIDS AND DERIVATIVES Preparation 1 2-Hydro\yimino(thien-2-yl)acetic acid (syn-isomer) A cooled solution of hydroxylamine hydrochloride (0.884 g) and sodium bicarbonate (L08 g) in water (20 ml.) was added to a cooled solution of thien- 2-yl-glyoxylic acid (2.0 g) and sodium bicarbonate (1.08 g) in water (20 ml.). After 2 days at 20 the solution was extracted with ether. cooled and acidified with concentrated hydrochloric acid. The resulting white solid was filtered off (0564 g) and the filtrate was then thoroughly extracted with ether. The combined extracts were washed with brine. dried and concentrated. The solid residue was triturated with benzene to give the oxime (syn-isomer) (1.05 g; l7l). m.p. l32,)\,,,,,, (ethanol) 284 nm (e 9.500), u

12 (Nujol) 2590 and 1706 (CO- H) and 1655 cm (C=N). 1-(DMSO-do). -25 to -0.5 (broad multiplet; N-OH and COOH). 2.36 (multiplet; thienyl C-S H) and 2.82 (multiplet'. thienyl 3 H and C-4 H).

Preparation 2 2-Dichloroacetoxyiminophenylacetic acid (syn-isomer) To a mixture of methylene chloride (45 ml) and dichloroacetyl chloride 10 ml) was added portionwise with stirring 2-hydroxyiminophenylacetic acid (synisomer) (5g in portions over about fifteen minutes. The reaction mixture became solid. and was stirred for one hour at room temperature alter the addition was complete. The reaction was diluted with petroleum (Imp. -l()-(0). filtered. and the solid washed several times with petroleum spirit to remove residual acid chloride. The solid was dried under vacuum. giving 2- dichloroacetoxyiminopbenylacetic acid (synisomer)(8.0g'. 969i mp 115. u,,,,, (CHBm) 3470 and l75l) (CO:H). I765 cm. (ester). 7 (CDCl values include 2.0 2.7 (multiplet; aromatic protons). 3.85 (singlet; CHCl;).

Preparation 3 2-Dichlortnicetoxyiminw(thicn-2-yl)acetic acid (synisomer) 2-Hydroxyimino-(thien 2-yl)acetic acid (syn-isomer) (38g) was added portionwise to a stirred solution 01 dichloroacetyl chloride ml.) in dry methylene chlo ride (350 ml.) at 20. Stirring was continued and after ca. 20 mins. White fibrous crystals formed. The suspension was stirred for 10 mins. longer and then filtered. The solid was washed with methylene chloride and petroleum to give the title acid (38.5 g.. 61%) )v,,,,, (EtOH) 262.5 (6 9.520). 291 nm (c 8.580).

Preparation 4 Z-Hydrox yimino-2( thien 3-yl)-acetic acids To a stirred solution of hydroxylamine hydrochloride (1.15 g) in methanol (15 mls) containing phenolphthalein (2 drops) was added a solution of sodium methoxide in methanol until a pink colour was obtained. After the addition of one crystal of hydroxylamine hydrochloride to discharge the pink colour the mixture was added to a solution of thien-3-yl-glyoxylic acid (2.0 g) in methanol (l0 mls) and refluxed for l hr. The methanol was evaporated to a small volume and the residue was partitioned between ethyl acetate and water. The aqueous layer was acidified to pH 1 with 2N- hydrochloric acid and extracted with ethyl acetate. After drying. the ethyl acetate was evaporated to yield a white solid consisting of isomeric 2-hydroxyimino-2- (thien-3-yl) acetic acids. h (ethanol) 254.5 nm (5 9.700). (CHE 3540 (OH unbonded). 3200 (OH bonded) and I750 cm. (COOH) 'r (DMSOd values include l.78 lmultiplet; thienyl protons (anti isomer)] 2.2 to b 2.7 (complex multiplet; thienyl protons).

Preparation 5 2-Dichloroacetoxyimino-2-(thien-3-yl)acetyl chloride (syn isomer) 2-Hydroxyimino-2-(thien-3-yl)acetic (1.03 g) mixture was added portionwise to a stirred solution of dichloroacetyl chloride (1.8 mls) in dry methylene chloride (l0 mls). After 30 mins. an excess of petrol (b.p. 40-60) was added to the reaction and the precipitated white solid was collected. washed with petrol (b.p. 40-60) and dried to yield 2-dichloroacetoxyimino-2-(thien-3-yl) acetic acid (syn isomer)( 1.6 gms. 94%) which was suspended in dry methylene chloride and ice-cooled. To the suspension was added dropwise a freshly prepared soltuion of phosphorus pentachlo ride 1 equiv.) in dry methylene chloride. When all components were in solution the solvent was removed in vacuo at low temperature and the residual oil was azeotroped with benzene to give 2-dichloroacetoxyimino-2-(thien-3-yl)acetyl chloride (syn) as a pale yellow oil which was used directly.

Preparation 6 2-Hydroxyimino-2-(fur-2-yl )acetic acids To a stirred solution of hydroxylamine hydrochloride 1.94 gms) in methanol (30 mls) containing phenolphthalein (2 drops) was added a solution of sodium methoxide in methanol until a pink colour was obtained. After the addition of one crystal of hydroxylamine hydrochloride to discharge the pink colour the mixture was added to a solution of fur-2ylglyoxylic acid (3.0 gm) in methanol (10 mls) and refluxed for 1 hr. The methanol was evaporated to a small volume and partitioned between ethyl acetate and water. The aqueous layer was acidified to pH 1 with ZNhydrochloric acid and extracted with ethyl acetate. After drying. the solvent was evaporated to yield a yellow solid which was collected. washed with petrol (b.p. 4060) and dried to yield an isomeric mixture of 2-hydroxyimino-2-( fur- 2-yl)acetic acids (2.7 gms. 81%) A (ethanol) 27 I .5 nm (6. H.400). v,,,,,., (CHBH) 3550 (OH). 1740 and 1705 cm? (CO H). T DMSO d) values include 2.62 [doublet .l 4 Hz. furyl(anti isomer) C 3-H]. 2.14 and 3.32 (complex signals. remaining furyl protons).

Preparation 7 2-Dichloroacetoxyimino-2-(fur-2-yl )acetyl chloride yn) 2-Hydroxyimino-2-(fur-2-yl)acetic acid (l.38g)mixture was added portionwise to an ice cold solution of dichloroacetyl chloride (2.8 mls.) in dry methylene chloride l4 mls). After mins stirring a white precipitate was formed which was collected. washed successively with cold dry methylene chloride and petrol (b.p. 40-60) and dried to yield Z-diehloroacetoxyimino-2-(fur-2-yl)acetic acid (syn-isomer) (1.5 g. 63%).

To an ice-cold stirred suspension of 2- dichloroacetoxyimino-2-(fur-2-yl)acetic acid l.5 g) in dry methylene chloride (70 mls) was added dropwise a freshly prepared solution of phosphorus pentachloride 1.0 equiv.) in methylene chloride. When solution was complete the solvent was evaporated at low temperature and the residual oil azeotroped with benzene to give 2-dichloroacetoxyimino-2(fur-Z-yl )acetyl chloride (syn-isomer) as a pale yellow oil which was used di rectly.

EXAMPLE 1 6B-( Z-Hydroxyimino-Z-p henylacetamido )-2,2-dimethylpenam-3a-carboxylic acid (syn-isomer) To a suspension of fifi-amino-2.Z-dimethylpenam-liacarboxylic acid (1.08 g) in chloroform ml) was added hexamethyldisilazane (2.49 g) and the mixture was heated under reflux for 2 hours, during which time solution occurred. The solvents were removed under reduced pressure. giving trimethylsilyl 2.2-dimethylfafi-trimethylsilylaminopenam-Rwearboxylate as a solid. The solid was dissolved in methylene chloride (20 ml) and propylene oxide 1.5 ml) added. The solution was cooled to 0 and treated dropwise with 2- dichloroacetoyyimino-2-phenylacetyl chloride (from 1.38 g. 2-dichloroacetoxyiminoQ-phenylacetic acid (syn'isomerH in methylene chloride (20 ml). over a period of 15 minutes with stirring. The solution was allowed to stir for 2 hours at room temperature. The solvents were evaporation under reduced pressure below 35. and the residue taken up in ethyl acetate. After addition of methanol (2 ml). the solution was extracted as rapidly as possible with ice-cold sodium bicarbonate solution and the combined UXII'UCIS washed with ethyl acetate and poured into a stirred mixture of ethyl acetate and 2N-h \drochloric acid. The organic phase was separated and combined with an ethyl acetate extraction of the aqueous phase. and the solution washed with water. dried and evaporated. The residue was taken up in ethyl acetate (5 ml) and added dropwise to petroleum (b.p. 40h(|; 200 ml). The precipitated solid was filtered and dried. giving M34 2- hydroxyimino-2-phenylacetamido )2.2-dimeth \lpe nam-3a-earboxylic acid (syrrisomerl (0.84 g; 46% (all, 2l0 (c 09. DMSO). MW. (EtOH) 251 nm (e 9.450). VIM, (Nujol) 3650 2300 (OH I. 3280 (NH). l758 (li-lactam). 1735 (CO H). lo50 and I510 cm ((TONH). 1 (DMSO-do) 0.5l (doublet. J 7 HI; NH). 2.2-2.7 (multiplet; aromatic protons l. 4.28 (quartet. .l 4 Hz and 7 H7: C-oH). 4.40 (doublet J 4 H7; (5H). 5.27 (singlet; 3 H). 8.3) and 8.48 (two singlets. methyl groups).

EXAMPLE 2 2.2-Dimethyl-6B-I Z-hydroxyiminot thien-2 yl)acetamidol-penam-3u-carboxylic acid (sy n-isomer) A solution of trimethylsilyl 2.2dimethyl-oB-trimethylsilylaminopenam-Krt-earboxylate [prepared from the corresponding amino-acid (0.864 g. )1 and propylene oxide (1 ml.) in dichloromethane (20 m.) a cooled to 0 and treated dropwise with a solution of 2-dichloroacetoxyimino( thien-2-yl )acetyl chloride synisomer'. [prepared from the Corresponding acid (0.753 g.)] in dichloromethane (l2 ml.). The resulting solution was maintained at 0 for l hour and was then evaporated to a small bulk and partitioned between ethylacetate and saturated sodium bicarbonate solution. The aqueous phase was washed with ethyl acetate. acidified with 2N-hydrochloric acid. and extracted with ethyl acetate. The dried organic solution was evaporated to a small bulk and added dropwise to stirred petroleum spirit (b.p. 40-60). The resulting precipitate was collected. washed. and dried to afford the title acid (0.337 g.. 24% based on amine). [or], 224 (c 0.57. DMSO). Mm. (pH 6 buffer) 288 nm (6 6.400). inf. 270 nm (c 5.800). u (Nujol). 2600 and [730 (CO H). l776 (B-lactam). 1670 and l526 cmf' (CONH). T (d -DMSO) values include 0.44 (d. J 6 Hz; NH). 2.45 and 2.9 (multiplets. aromatic protons). 5.78 (s; C-3H). 8.46 and 8.55 (2 s; CH groups).

EXAMPLE 3 2.2-Dimethyl-6B-[2-hydroxyimino-2-(thien-3- yl)acetamide]-penam-3a-carboxylic acid (syn-isomer) To a cold solution of trimethylsilyl 2.2-dimethyl-6B trimethyl silylamino-penam-3 a-carboxylate [made from bfi-aminol.Z-dimethyl-penam-Tia-carboxylic acid (0.43 g) and hexamethyldisilazane (ll mls.)] and propylene oxide (053 mls) in dry methylene chloride l0 mls) was added dropwise with stirring a solution of Z-dichloroacetoxyimino-lthien-3-yl) acetyl chloride (sy n-isomer) (ll mmoles) in dry methylene chloride. After 1 hr. at C the reaction was evaporated and the residue dissolved in ethyl acetate. The ethyl acetate solution was extracted with ice'cold saturated sodium bicarbonate solution. The combined aqueous phases were acidified to pH 1 with ZN-hydrochloric acid and extracted with ethyl acetate. After drying the extracts were evaporated to a small volume which on adding to an excess of petrol lbp. 40-60) gave a white solid which was filtered and dried to yield the title com pound (0.33 g. 44% [61] +220 (c 09. dioxan). A (pH 6 buffer) 256 nm [e 9.]00). v,,,,, (Nujol) 3300 (NH). l7? lB-lactam). 1740 and 2600 (COOH) and 1660 and i526 cm. (CONH) 1' (DMSO Llli includestL-lb (d1J 8 H2; NH) 2.30. 2.35. 2.59 )m: thienyl protons). 5.66 (s; C-BH). and 8.35 and 8.44 (2s; CH; groups).

EXAMPLE 4 Z.Z-Dimethyl-oB-[ Z-hydroxyimino( fur-2- yl )acetamido l-penam-3a-c arboxylic acid (syn-isomer) A solution of trimethylsilyl ZQ-dimethyl-bB-trimethylsilylaminopenam-3a-carboxylate [prepared from the corresponding amino-acid (0.648 g.)] and propylene oxide (075 ml) in dichloromethane (15 ml) was cooled to 0 and treated dropwise with a solution of Z-dichloroacetoxyimino(fur-2-yl )acetyl chloride [synisomer; prepared from the corresponding acid (0.579 g.)] in dichloromethane (l2 ml.). The resulting solution was maintained at 0 for l hour and was then evaporated to small volume and partitioned between ethylacetate and saturated sodium bicarbonate soltuion. The aqueous phase was washed with ethyl acetate. acidified with ZN-hydrochloric acid and extracted with ethyl acetate. The dried organic solution was evaporated to a small bulk and added dropwise to stirred petroleum spirit (b.p. 40-60). The resulting precipitate was collected. washed and dried to give the title acid (0.56 g.. 55% based on amine), [01],, 243 (c 0.93. DMSO). (pH 6 buffer) 275 nm (c 9.600),

11 (Nujol). I778 (filactam). l740 and 2600 (CO H). 1670 and l52 cm? (CONH r (d -DMSO) values include 044 (d. J 6 H2; NH) 218 and 3.34 (multiplets: aromatic protons). 5.70 is; C-BH). 8.36 and 8.46 (2s; CH groups).

We claim:

I. A compound selected from the group consisting of a penicillin antibiotic of the formula R c CONH COOH wherein R is furyl or furyl substituted by chloro. bromo. hydroxy. lower alkyl. nitro. amino. lower alkylamino. di(lower alkyl)amino. lower alkanoyl, lower alkanoylamido. lower alkoxy. lower alkylthio or carbamoyl; and R" is hydrogen or 3oz-carboxylic acid (syn isomer).

UNITED STATES PATENT m 1 .1, CERTIFICATE OF CORRECTION Patent No. 3, 9 Dated December 16, 1975 I vent MICHAEL GREGSON et al.

It is certified that error appears in the above-identified patent and that said Letters Patent are hereby Corrected as shown below:

On the title page, between lines 13 and 14, there should appear the following:

- Foreign Application Priority Data January 29, 1971 Great Britain 3531/71 October 1, 1971 Great Britain .45885/71 January 14, 1972 Great Britain .cognated complete specification of 3531/71 and 45885/71 July 7, 1972 Great Britain. 32004/72-.

Signed and Scaled this thirtieth D a)/ of March 1976 [SEAL] Attest.

RUTH C. MASON C. MARSHALL DANN Alrcsling Officer ('ummissiunvr nj'lurenrs and Trademarks 

1. A COMPOUND SELECTED FROM THE GROUP CONSISTING OF A PENICILLIN ANTIBIOTIC OF THE FORMULA
 2. The compound of claim 1 which is 6 Beta -(2-hydroxyimino-2-(fur-2-yl)acetamido)-2,2-dimethylpenam-3 Alpha -carboxylic acid (syn isomer). 